Fluorescence Measurements of [Ca2+]I after shear stress of RBL cells

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Fluorescence Measurements of [Ca2+]I after shear stress of RBL cells Effects of Shear Stress on Intracellular Calcium Change and Histamine Release in Rat Basophilic Leukemia (RBL-2H3) Cells Fei Wei, WenzhongYang, Luwei Zhou Phys. Dept., Fudan Univ., Shanghai, People’s Republic of China Abstract: A mast cell RBL-2H3 was used to study cellular responses to shear stress generated by a rotating rotor in a cell dish. The intracellular calcium was studied by confocal fluorescence microscopy with Fluo-3/AM staining and the released histamine was measured with a fluorescence spectrometer using o-phthalaldehyde (OPA ) staining. An elevation of [Ca2+] occurred immediately after the shear stress, followed by histamine release. Furthermore, Ruthenium red, a transient receptor potential vanilloid (TRPV) inhibitor, could effectively block the shear stress–induced histamine release, suggesting that TRPV membrane proteins are the likely targets of the shear stress. Fluorescence Measurements of [Ca2+]I after shear stress of RBL cells 1.Stained with fluo-3/AM 2.Confocal fluorescence microscopy 3.Ca2+ fluorescence intensities in a cell in the Ca 2+free saline: η-viscosity ω-rotation speed Shear  [Ca2+ ]i Histamine detecting method TRPV4 channel OPA邻苯二甲醛+组胺 Cell viability Shear  histamine Shear  TRPV Ca2+ & histamine Conclusion : The change of [Ca2+ ]i and then histamine release can be induced by shear stress in RBL-2H3 cells . We speculate the TRPV4 channel involves in the response of RBL-2H3 to the physical stimuli. We would study the response of RBL cells to reciprocating force. Reference: [1] Wenzhong Yang et al. Effects of Shear Stress on Intracellular Calcium Change and Histamine Release in Rat Basophilic Leukemia (RBL-2H3) Cells, Journal of Environmental Pathology, Toxicology and Oncology, 28(3):223–230 (2009) [2] Wen-Zhong Yang et al. Effects of Low Power Laser Irradiation on Intracellular Calcium and Histamine Release in RBL-2H3 Mast Cells, Photochemistry and Photobiology, 2007, 83: 979–984